Treatment of osteosarcoma by targeting transportation of pHSP70-shPLK1/DOX compounds with bacterial magnetosomes

doi:10.3969/j.issn.2095-4344.2017.18.019

Abstract

Abstract:

BACKGROUND: In an additional changing magnetic field, magnetosomes can invade the tumor cells and inhibit their proliferation.

OBJECTIVE: To study the effect of targeted delivery of pHSP70-shPLK1/DOX compounds with magnetosome of magnetotactic bacteria on the treatment of osteosarcoma.

METHODS: Human osteosarcoma cell U2OS was divided into four groups: group A: magnetosomes of magnetotactic bacteria+pHSP70-shPLK1/DOX complex; group B: magnetosomes of magnetotactic bacteria; group C: magnetosomes of magnetotactic bacteria+pHSP70-shPLK1/DOX complex, combined with magnetic intervention; group D: magnetosomes of magnetotactic bacteria, combined with magnetic intervention. After 12, 24, 48 and 72 hours of culture, the uptake rate of osteosarcoma cells was observed; cell cycle was evaluated by flow cytometry; PLK1 mRNA and protein expression levels were detected by RT-PCR and western blot, respectively; cell proliferation was explored by MTT assay; cell adhesion and invasion were checked by adhesion assay and Transwell chamber assay, respectively; and the apoptosis rate was measured by apoptosis detection Kit.

RESULTS AND CONCLUSION: (1) After culture of 12 and 24 hours, the uptake rate was the highest in group D; meanwhile, the uptake rate was the highest in group B after 48 and 72 hours of culture. (2) The ratio of G₂/M phase decreased and the ratio of G₀/G₁ increased gradually in group A, group C and group D; the G₂/M phase ratio of C group was the lowest at each time point, followed by group D, and that of group A was the highest; and the G₂/M phase ratio increased gradually in group B. (3) Following 12 and 24 hours of culture, the expression of PLK1 in group B and group D was higher than that in group A and group C (P < 0.05); in addition, the expression of PLK1 in group B was higher than that in the other three groups after 48 and 72 hours of culture (P < 0.05), and the PLK1 level in group D was higher than that in group A and group C (P < 0.05). (4) The proliferation, adhesion and invasion ability in group B were the highest at different time points, but the apoptosis rate was the lowest. Furthermore, the proliferation, adhesion and invasion abilities in group C were the lowest at different time points, and the apoptosis rate was the highest. To conclude, our experimental results show that the targeted delivery of pHSP70-shPLK1/DOX complex by magnetosomes of magnetotactic bacteria can enhance the apoptosis of osteosarcoma cells and inhibit their proliferation and invasion.

中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程

Key words: Magnetosomes, Osteosarcoma, Cell Proliferation, Apoptosis, Tissue Engineering

CLC Number:

R318

Yu Shui-sheng, Cheng Li, Xu Xin-zhong, Ke You-qun, Jing Jue-hua.

Treatment of osteosarcoma by targeting transportation of pHSP70-shPLK1/DOX compounds with bacterial magnetosomes [J]. Chinese Journal of Tissue Engineering Research, 2017, 21(18): 2906-2912.

Figures/Tables 6

References

[1]Luetke A,Meyers PA,Lewis I,et al.Osteosarcoma treatment-Where do we stand? A state of the art review. Cancer Treatt Rev.2013;40(4):523-532.

[2]Tacar O,Sriamornsak P,Dass CR.Doxorubicin: an update on anticancer molecular action, toxicity and novel drug delivery systems.J Pharm Pharmacol.2013;65(2):157-170.

[3]Seow HF,Yip WK,Fifis T.Advances in targeted and immunobased therapies for colorectal cancer in the genomic era.Onco Targets Ther.2016;9:1899-1920.

[4]Melero I,Berman DM,Aznar MA,et al.Evolving synergistic combinations of targeted immunotherapies to combat cancer. Nat Rev Cancer.2015;15(8):457-472.

[5]Galluzzi L,Buque A,Kepp O,et al.Immunological Effects of Conventional Chemotherapy and Targeted Anticancer Agents. Cancer Cell.2015;28(6):690-714.

[6]Chou YS,Yen CC,Chen WM,et al.Cytotoxic mechanism of PLK1 inhibitor GSK461364 against osteosarcoma: Mitotic arrest,apoptosis,cellular senescence,and synergistic effect with paclitaxel.Int J Oncol.2016;48(3):1187-1194.

[7]Ma H,He C,Cheng Y,et al.PLK1shRNA and doxorubicin co-loaded thermosensitive PLGA-PEG-PLGA hydrogels for osteosarcoma treatment.Biomaterials. 2014;35(30): 8723-8734.

[8]Gjertsen BT,Schoffski P.Discovery and development of the Polo-like kinase inhibitor volasertib in cancer therapy. Leukemia.2015;29(1):11-19.

[9]Cheng L,Wang C,Jing J. Polo-like kinase 1 as a potential therapeutic target for osteosarcoma.Curr Pharm Des.2015; 21(10):1347-1350.

[10]Zhao L,Jiang BO,Wang D,et al.Triptolide reduces the viability of osteosarcoma cells by reducing MKP-1 and Hsp70 expression.Exp Ther Med.2016;11(5):2005-2010.

[11]姚琴,赵慧毅,谢柏臻.Ezrin shRNA联合HSP70对骨肉瘤细胞凋亡和增殖的影响[J].中华骨科杂志,2015, 35(2):165-173.

[12]Mathuriya AS.Magnetotactic bacteria: nanodrivers of the future.Crit RevBiotechnol. 2015;22(6):1-15.

[13]Goldhawk DE,Gelman N,Thompson RT,et al.Forming Magnetosome-Like Nanoparticles in Mammalian Cells for Molecular MRI[M]//Design and Applications of Nanoparticles in Biomedical Imaging.Springer International Publishing, 2017:187-203.

[14]张薇薇,许乙凯,刘瑞源.基于趋磁细菌磁小体的新型主动靶向线粒体的MR双功能探针制备[J].中华放射学杂志, 2014,48(6): 509-511.

[15]Chen CY,Chen CF,Yong Y,et al.Construction of a microrobot system using magnetotactic bacteria for the separation of Staphylococcus aureus.Biomed Microdevices. 2014;16(5): 761-770.

[16]Alphandéry E,Guyot F,Chebbi I.Preparation of chains of magnetosomes, isolated from Magnetospirillum magneticum, strain AMB-1 magnetotactic bacteria, yielding efficient treatment of tumors using magnetic hyperthermia.Int J Pharm. 2012;434(1-2):444-452.

[17]Yuan-Gang L,Qing-Lei D,Shi-Bin W,et al.Preparation and in vitro antitumor effects of cytosine arabinoside-loaded genipin-poly-l-glutamic acid-modified bacterial magnetosomes. Int J Nanomedicine. 2015;10:1387-1397.

[18]Isakoff MS,Bielack SS,Meltzer P,et al.Osteosarcoma: Current Treatment and a Collaborative Pathway to Success.J Clin Oncol.2015;33(27):3029-3035.

[19]Gill J,Ahluwalia MK,Geller D,et al.New targets and approaches in osteosarcoma.Pharmacol Ther. 2013;137(1): 89-99.

[20]Teagle AR,Birchall JC,Hargest R.Gene Therapy for Pyoderma Gangrenosum: Optimal Transfection Conditions and Effect of Drugs on Gene Delivery in the HaCaT Cell Line Using Cationic Liposomes.Skin Pharmacol Physiol. 2016;29(3): 119-129.

[21]Lohße A,Ullrich S,Katzmann E,et al.Functional Analysis of the Magnetosome Island in Magnetospirillum gryphiswaldense: The mamAB Operon Is Sufficient for Magnetite Biomineralization. Plos One.2011;6(10):e25561.

[22]Alphandéry E,Chebbi I,Guyot F,et al.Use of bacterial magnetosomes in the magnetic hyperthermia treatment of tumours: A review.Int J Hyperthermia.2013;29(8):801.

[23]Hervault A,Dunn AE,Lim M,et al.Doxorubicin loaded dual pH-and thermo-responsive magnetic nanocarrier for combined magnetic hyperthermia and targeted controlled drug delivery applications. Nanoscale.2016;8(24):12152.

[24]Sun J,Li Y,Liang XJ,et al.Bacterial magnetosome: a novel biogenetic magnetic targeted drug carrier with potential multifunctions.J Nanomater. 2011;2011(2011):469031- 469043.

[25]王熙,耿圆圆,侯蔷,等.细菌磁小体作为新型非病毒载体用于肿瘤基因治疗的研究进展[J].肿瘤学杂志, 2016,22(4):322-326.

[26]尹燕鹰,顾宁,洪敏,等.靶向磁性热疗对小鼠结肠癌皮下移植模型作用的观察[J].中华肿瘤防治杂志,2013, 20(4):249-253.

[27]Li C,Ke Y,Yu S,et al.Co-delivery of doxorubicin and recombinant plasmid pHSP70-Plk1-shRNA by bacterial magnetosomes for osteosarcoma therapy.Int J Nanomedicine. 2016;11:5277-5286.

[28]Sero V,Tavanti E,Vella S,et al.Targeting polo-like kinase 1 by NMS-P937 in osteosarcoma cell lines inhibits tumor cell growth and partially overcomes drug resistance.Invest New Drugs.2014; 32(6):1167-1180.

[29]Otsu H,Iimori M,Ando K,et al.Gastric Cancer Patients with High PLK1 Expression and DNA Aneuploidy Correlate with Poor Prognosis.Oncology.2016;2(6):15-16.

[30]Fischer M,Quaas M,Nickel A,et al.Indirect p53-dependent transcriptional repression of Survivin, CDC25C, and PLK1 genes requires the cyclin-dependent kinase inhibitor p21/CDKN1A and CDE/CHR promoter sites binding the DREAM complex.Oncotarget.2015;6(39):41402-41417.

[31]Ogawa R,Morii A,Watanabe A,et al.Regulation of gene expression in human prostate cancer cells with artificially constructed promoters that are activated in response to ultrasound stimulation.Ultrason Sonochem.2012;20(1): 460-467.

[32]Bhayani KR,Rajwade JM,Paknikar KM.Radio frequency induced hyperthermia mediated by dextran stabilized LSMO nanoparticles: in vitro evaluation of heat shock protein response. Nanotechnology.2013;24(1):015102.

[33]Yamaguchi M,Ito A,Okamoto N,et al.Heat-inducible transgene expression system incorporating a positive feedback loop of transcriptional amplification for hyperthermia-induced gene therapy.J Biosci Bioeng.2012;114(4):460-465.

[34]Yamaguchi M,Ito A,Ono A,et al.Heat-inducible gene expression system by applying alternating magnetic field to magnetic nanoparticles.ACS Synth Biol 2014;3(5):273-279.

[35]Tantos A,Kalmar L,Tompa P.The role of structural disorder in cell cycle regulation, related clinical proteomics, disease development and drug targeting.Expert Rev Proteomics.2015; 12(3):221-233.

[36]Liu Y,Bi T,Dai W,et al.Lupeol Induces Apoptosis and Cell Cycle Arrest of Human Osteosarcoma Cells Through PI3K/AKT/mTOR Pathway.TechnolCancer Res Treat.2016; 15(6):NP16.